FIG 5 .

S. aureus induces autophagy in keratinocytes. (A) Confocal images of galectin-8 (red)- or LC3 (red)-labeled sections of SCID-hu grafts from mice infected with WT S. aureus (SA) or agr mutant (green). Representative images from two separate experiments are shown. (B) Electron micrograph of cells infected with agr null mutant demonstrating the double membrane surrounding staphylococci and adjacent mitochondria. (C) Intracellular recovery of WT S. aureus USA300 and agr mutant from HaCaT and HEKn cells treated with wortmannin (24-h infection) or 3-MA (2-h infection). Data shown represent the means plus standard errors of sextuplicate samples; at least two independent experiments were done, and the results of a representative experiment are shown. The P values in the graphs compare the value for the strain shown to the value for no inhibitor by Student’s t test. (D) Immunoblot of WT S. aureus- or agr mutant-infected HaCaT cells compared to a PBS control detecting galectin-8 and LC3II. (E) Flow cytometric analysis of autophagy in HaCaT cells exposed to WT USA300 or agr null mutant by Cyto-ID (LC3) staining in the presence of wortmannin, 3-MA (negative control), and chloroquine (positive control) to increase autophagosome formation.